Invasin of Yersinia pseudotuberculosis is not a polyclonal activator of mouse B lymphocytes

It is known that the invasin molecule of Yersinia pseudotuberculosis stimulates human peripheral B cells in vitro. In this work we evaluated the in vivo role of invasin as polyclonal activator of B lymphocytes in the mouse experimental model, by comparing strains of Y. pseudotuberculosis expressing invasin and isogenic inv mutants. Swiss mice were infected intravenously with two strains expressing invasin (YpIII pIB1 and an isogenic virulence plasmid-cured strain, YpIII) and with two invasin mutant strains (Yp100 pIB1 and Yp100, plasmid-cured). Spleen cells were sampled on days 7, 14, 21 and 28 after infection. Immunoglobulin (Ig)-secreting spleen cells were detected by protein A plaque assay and specific antibodies were detected in sera by ELISA. The virulent strain YPIII pIB1 (wild type) did not provoke polyclonal activation of B lymphocytes in vivo. In general, fewer Ig-secreting spleen cells of all isotypes were found in the infected animals than in the control animals. Specific IgG antibodies were detected in the sera of animals infected with all strains. The peak response occurred on the 21 st day post-infection, and the Yp100 strain provoked the highest level of these antibodies. We concluded that invasin is not a polyclonal activator of murine B cells.

Medium-term tongue carcinogenesis assays: A comparative study between 4-nitroquinoline 1-oxide (4NQO)-induced rat and dimethylbenzanthracene (DMBA)-induced hamster carcinogenesis

The most used animal models in oral cancer research are the hamster treated by dimethylbenzanthracene (DMBA), and the rat treated by 4-nitroquinoline 1-oxide (4NQO). The purpose of this study was to compare the DMBA-induced hamster tongue carcinogenesis and 4NQO-induced rat tongue carcinogenesis by means of morphological analysis. Male Wistar rats were distributed into three groups of ten animals each and treated with 50 ppm 4NQO solution by drinking water for 4, 12 or 20 weeks. A total of 18 Syrian golden hamsters were submitted to 0.5% DMBA (dissolved in acetone) topical application three times/week for 4, 12 and 20 weeks. The primary histopathological change i.e., hyperplasia and hyperkeratosis, was evidenced after 4 weeks treatment with DMBA. Regarding 12 weeks treatment, 4NQO and DMBA were able to induce morphological changes as depicted by hyperplasia and dysplasia. At 20 weeks, squamous cell carcinoma was found in the majority of animals for both carcinogens used. Taken together, our results suggest that the hamster experimental model disclosed aspects related with tongue carcinogenesis in lesser time than rats. Probably, such discrepancies depend strongly on route of administration and the susceptibility with respect to animal species. © 2006 Elsevier GmbH. All rights reserved.

Immune response to Mycobacterium bovis-an5 infection in genetically selected mice (selection IV-A)

Mice genetically selected for high (H) and low (L) antibody production (Selection IV-A) were used as murine experimental model. The aim of the present work was to evaluate the macrophagic activity and to characterize the immune response in Mycobacterium bovis-AN5 infected mice (3×10 7 bacteria). The response profile previously observed in such strains was not similar to that obtained during M. bovis infection; however, it corroborated works carried out using Selection I, which is very similar to Selection IV-A regarding infection by M. tuberculosis and Bacillus Calmette-Guérin (BCG). Considering bacterial recovery, LIV-A mice showed higher control of the infectious process in the lungs than in the spleen, whereas HIV-A mice presented more resistance in the spleen. With respect to macrophagic activity, hydrogen peroxide (H2O 2) was probably not involved in the infection control since there was an inhibition in the production of this metabolite. Nitric oxide (NO) and TNF-α production seemed to be important in the control of bacterial replication and varied according to the strain, period and organ. Evaluation of the antibody production indicated that the multi-specific effect commonly observed in these strains was not the same in the response to M. bovis. Antibody concentrations were higher in LIV-A than in HIV-A mice at the beginning of the infection, being similar afterwards. Such data were compared with delayed-type hypersensitivity (DTH), which was more intense in HIV-A than in LIV-A mice, indicating that antibody production is independent of the capability to trigger DTH reactions and that cellular and humoral responses to M. bovis antigens show a polygenic control and an independent quantitative genetic regulation. Differences were observed among organs and metabolites, suggesting that different mechanisms play an important role in this infection in natural heterogeneous populations, indicating that NO, TNF-α and Th1 cytokines are involved in the infection control.

Utilização do AutoCAD 2004 para quantificação de pesquisas usando fotomicrografias eletrônicas

Objective: To determine if the AutoCAD 2004 program is available to be used in quantitative research on electron microscopy. Methods: A demonstrative of the AutoCAD2004 was done in an experimental model of diabetes induced in rats. The animals were divided by lot in control group animals (GC-10 animals) and diabetic group (GD - 10 animals), evaluated 1 month after the diabetes induction.The eyes were removed after sacrifice and prepared to electron microscopy. The retina was photographed and the AutoCAD2004 program was used to measure the basal membrane of retina vessels. The results were submitted to statistical analysis. Results:The AutoCAD2004 was effective to obtain quantitative measures from the basal membrane of retina vessels and there were no difference between the studied groups. Conclusion: The AutoCAD2004 was effective, safe and easy to apply to quantify biological parameters, which allowed suggest it to quantitative research.

Microwave fixation in rat fetuses tissues: Histological and immunohistochemical analysis

The aim of this study was to show the microwaves action in fixation of rat fetuses, dermal and cartilaginous tissues, using histological and immunohistochemistry methods for analysis. It was possible to conclude in this study using the rat as experimental model that the two methods for antibody retrieval, presented an excellent ways for the use of Ki67 antibody in the immunohistochemical analysis.

Responses of blood lactate concentration in aerobic and anaerobic training protocols at different swimming exercise intensities in rats

A swimming periodized experimental training model in rats in which different training protocols (TP) were classified in aerobic (A) and anaerobic (AN) intensity levels. The purpose of the present study was to verify if the classification of the TP used in the periodized training experimental model presented the blood lactate concentration [La] response adequate to the aerobic and anaerobic intensities levels. Twenty three male Wistar rats were divided into three groups. Two groups of swimming training (continuous, CT, n = 7, and periodized training, PET, n = 7) rats were evaluated during 5 weeks in eight different TP (TP-1 to TP-8) through the analysis of the [La] response. The third group was the sedentary control (SC, n = 9). The TP were classified in five intensity levels, three aerobic (A-1, A-2, A-3) and two anaerobic (AN-1, AN-2). Analysis of variance (ANOVA one-way, P<0.05) indicated significant differences in the [La] among the TP and among the five intensity levels. All TP of the A-2 and A-3 intensity levels differed from the A-1 and AN-1. The A-1 and AN-1 also differed among them. These findings demonstrate that the TP were classified properly at different levels of aerobic and anaerobic intensities, as based on the [La] response in a way similar to that of high performance swimming with humans. The results offer new perspectives for the study of exercise training in swimming rats at different levels intensity for performance or for health.

Soybean glycinin (11S) increases HDL-cholesterol in hypercholesterolemic rats

Purpose: The purpose of this paper is to determine the effects of isolated soy glycinin (11S) on lipid metabolism in animals subjected to a hypercholesterolemic diet. Design/methodology/approach: Male Wistar rats were kept in individual cages under appropriate conditions. The animals were divided into three groups (n=9): normal diet (STD) given a diet containing casein as protein source, recommended in AIN-93M; hypercholesterolemic (HC) fed a normal diet with 1 per cent cholesterol and 0.5 per cent cholic acid; and hypercholesterolemic+glycinin (HC+11S), fed a hypercholesterolemic diet, plus 11S soy protein (300 mg/kg/day), dissolved in saline and administered by gavage. After 28 days, the animals were sacrificed and blood and liver removed for biochemical analysis of total cholesterol (TC), HDL-cholesterol (HDL-C) and triglycerides (TG) in the plasma, hepatic TC and TG. Findings: A single daily dose of glycinin given to the hypercholesterolemic group demonstrated its functional role, particularly in raising HDL-C and reducing triglycerides in the liver. Originality/value: This study demonstrates the action of the 11S globulin in soybean as a serum lipid lowering agent, in addition to its nutritional properties, especially in raising the HDL-C. © Emerald Group Publishing Limited.

Implant osseointegration in circumferential bone defects treated with latex-derived proteins or autogenous bone in dog's mandible

Background: In sites with diminished bone volume, the osseointegration of dental implants can be compromised. Innovative biomaterials have been developed to aid successful osseointegration outcomes. Purpose: The aim of this study was to evaluate the osteogenic potential of angiogenic latex proteins for improved bone formation and osseointegration of dental implants. Materials and Methods: Ten dogs were submitted to bilateral circumferential defects (5.0×6.3mm) in the mandible. Dental implant (3.3×10.0mm, TiUnite MK3™, Nobel Biocare AB, Göteborg, Sweden) was installed in the center of the defects. The gap was filled either with coagulum (Cg), autogenous bone graft (BG), or latex angiogenic proteins pool (LPP). Five animals were sacrificed after 4 weeks and 12 weeks, respectively. Implant stability was evaluated using resonance frequency analysis (Osstell Mentor™, Osstell AB, Göteborg, Sweden), and bone formation was analyzed by histological and histometric analysis. Results: LPP showed bone regeneration similar to BG and Cg at 4 weeks and 12 weeks, respectively (p≥.05). Bone formation, osseointegration, and implant stability improved significantly from 4 to 12 weeks (p≤.05). Conclusion: Based on methodological limitations of this study, Cg alone delivers higher bone formation in the defect as compared with BG at 12 weeks; compared with Cg and BG, the treatment with LPP exhibits no advantage in terms of osteogenic potential in this experimental model, although overall osseointegration was not affected by the treatments employed in this study. © 2009 Wiley Periodicals, Inc.

Estudo da segurança de cosméticos: Presente e futuro

Cosmetics have been used since ancient times and, recently, their consumption has increased greatly in many countries, including brazil, which is the third largest consumer market in the world. Thus, concern for the safety and efficacy of these products should be heightened, even though these products are rarely related to adverse reactions that damage the health. Brazilian law requires manufacturers to subject their products to safety testing, to assess the possible reactions that could be caused by them (irritation, sensitization, systemic effects). To this end, in general, animals have been used as the experimental model, but this practice is being increasingly controlled, so that the scientific community is looking for alternative tests that do not require experimental in vivo models. Thus, this review aims to describe the main biological assays used to assess the safety of cosmetics, as well as in vitro assays that can replace them.

Photodynamic therapy for the treatment of induced mammary tumor in rats

The objective of this work was to evaluate photodynamic therapy (PDT) by using a hematoporphyrin derivative as a photosensitizer and light-emitting diodes (LEDs) as light source in induced mammary tumors of Sprague-Dawley (SD) rats. Twenty SD rats with mammary tumors induced by DMBA were used. Animals were divided into four groups: control (G1), PDT only (G2), surgical removal of tumor (G3), and submitted to PDT immediately after surgical removal of tumor (G4). Tumors were measured over 6 weeks. Lesions and surgical were LEDs lighted up (200 J/cm2 dose). The light distribution in vivo study used two additional animals without mammary tumors. In the control group, the average growth of tumor diameter was approximately 0.40 cm/week. While for PDT group, a growth of less than 0.15 cm/week was observed, suggesting significant delay in tumor growth. Therefore, only partial irradiation of the tumors occurred with a reduction in development, but without elimination. Animals in G4 had no tumor recurrence during the 12 weeks, after chemical induction, when compared with G3 animals that showed 60 % recurrence rate after 12 weeks of chemical induction. PDT used in the experimental model of mammary tumor as a single therapy was effective in reducing tumor development, so the surgery associated with PDT is a safe and efficient destruction of residual tumor, preventing recurrence of the tumor. © 2012 Springer-Verlag London Ltd.

Comparisons between Bio-Oss® and Straumann® Bone Ceramic in immediate and staged implant placement in dogs mandible bone defects

Objective: To compare immediate and staged approach implant placement in circumferential defects treated with deproteinized bovine bone mineral (DBBM); hidroxyapatite/tricalcium phosphate (HA/TP); autogenous bone (Ab); and coagulum (Cg); upon implant stability, osseointegration and alveolar crest maintenance. Materials and methods: Six dogs underwent extractions of lower premolars, bilaterally. Twelve weeks later four bone defects (6 mm wide/4 mm long) were drilled at one side and randomly filled with DBBM; HA/TP; Ab; and Cg, respectively, and left to heal (staged approach). Eight weeks later one implant (Osseospeed™, AstraTech) was placed in experimental sites. At the same session four defects were drilled on contra-lateral side and implants were inserted immediately after biomaterials grafting (immediate approach). Animals were euthanized 8 weeks later. Implant stability was measured by resonance frequency analysis (RFA) at installation and after sacrifice. Ground sections were prepared for bone contact (BIC); bone area (BA); distance implant shoulder-bone crest (IS-C); distance implant shoulder first bone contact (IS-B); and areas occupied by soft tissue. Results: The BA and BIC were superior in the staged approach. The Cg exhibited higher BIC and BA as compared with other materials at the total implant body (P = 0.004 and 0.012, respectively). The DBBM, HA/TP and Ab groups rendered similar BA and BIC. The immediate approach resulted in less crest resorption compared to staged approach. The biomaterials did not affect the IS-C and IS-B measurements. Particles area tended to be higher in DBBM group than HA/TP (P = 0.15), while soft tissue infiltrate was higher in DBBM group when used in the immediate approach (P = 0.04). The RFA indicated gain in stability in the staged approach (P = 0.002). The correlation test between RFA vs. BIC and BA demonstrated inferior stability for DBBM group in immediate approach (P = 0.01). Conclusions: Implants placed in healed defects resulted in better stability as a consequence of higher BIC and BA. The Cg alone rendered increased BIC compared to other materials in both approaches. Immediate approach should be preferable to staged approach in terms of alveolar crest maintenance. The BIC and BA values did not vary between micro and macro-threads in this experimental model. Implants installed in sites filled with DBBM in immediate approach were less stable. © 2011 John Wiley & Sons A/S.

Motor Recovery and Synaptic Preservation after Ventral Root Avulsion and Repair with a Fibrin Sealant Derived from Snake Venom

Background:Ventral root avulsion is an experimental model of proximal axonal injury at the central/peripheral nervous system interface that results in paralysis and poor clinical outcome after restorative surgery. Root reimplantation may decrease neuronal degeneration in such cases. We describe the use of a snake venom-derived fibrin sealant during surgical reconnection of avulsed roots at the spinal cord surface. The present work investigates the effects of this fibrin sealant on functional recovery, neuronal survival, synaptic plasticity, and glial reaction in the spinal motoneuron microenvironment after ventral root reimplantation.Methodology/Principal Findings:Female Lewis rats (7 weeks old) were subjected to VRA and root replantation. The animals were divided into two groups: 1) avulsion only and 2) replanted roots with fibrin sealant derived from snake venom. Post-surgical motor performance was evaluated using the CatWalk system twice a week for 12 weeks. The rats were sacrificed 12 weeks after surgery, and their lumbar intumescences were processed for motoneuron counting and immunohistochemistry (GFAP, Iba-1 and synaptophysin antisera). Array based qRT-PCR was used to evaluate gene regulation of several neurotrophic factors and receptors as well as inflammatory related molecules. The results indicated that the root reimplantation with fibrin sealant enhanced motor recovery, preserved the synaptic covering of the motoneurons and improved neuronal survival. The replanted group did not show significant changes in microglial response compared to VRA-only. However, the astroglial reaction was significantly reduced in this group.Conclusions/Significance:In conclusion, the present data suggest that the repair of avulsed roots with snake venom fibrin glue at the exact point of detachment results in neuroprotection and preservation of the synaptic network at the microenvironment of the lesioned motoneurons. Also such procedure reduced the astroglial reaction and increased mRNA levels to neurotrophins and anti-inflammatory cytokines that may in turn, contribute to improving recovery of motor function. © 2013 Barbizan et al.

Slimmer or Fertile? Pharmacological Mechanisms Involved in Reduced Sperm Quality and Fertility in Rats Exposed to the Anorexigen Sibutramine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação leucocitária de cães com linfoma submetidos ao protocolo de Madison-Wisconsin pela técnica convencional e citometria de fluxo

Lymphoma is the most common hematopoietic tumor in dogs and one of the malignant tumors with higher occurrence in this species. It is a great experimental model due to its resemblance with the non-Hodgkin lymphoma in humans. Considering the importance of the overall changes that result from to this kind of neoplastic tumor and those due to polichemotherapy this study aimed to evaluate the absolute leukon count and the total count of CD45+ cells in the blood of 25 dogs with lymphoma. Findings were crosschecked since diagnose and then once weekly during the first eight sessions of the Madison-Wisconsin chemotherapic protocol. Total granulocyte, lymphocyte and monocyte counts obtained from a conventional automatic counter and by flow cytometry were compared. Results did not reveal statistically significant changes between the two techniques.